Page 1
REVIEW ARTICLE
Extraction methods of butterfly pea (Clitoria ternatea) flowerand biological activities of its phytochemicals
Ethel Jeyaseela Jeyaraj1 · Yau Yan Lim1 · Wee Sim Choo1
Revised: 17 July 2020 / Accepted: 18 August 2020 / Published online: 1 September 2020
© Association of Food Scientists & Technologists (India) 2020
Abstract Clitoria ternatea or commonly known as ‘But-
terfly pea’ has been used traditionally in Ayurvedic medi-
cine in which various parts of the plants are used to treat
health issues such as indigestion, constipation, arthritis,
skin diseases, liver and intestinal problems. The flowers of
C. ternatea are used worldwide as ornamental flowers and
traditionally used as a food colorant. This paper reviews
the recent advances in the extraction and biological activ-
ities of phytochemicals from C. ternatea flowers. The
application of maceration or ultrasound assisted extraction
greatly increased the yield (16–247% of increase) of phy-
tochemicals from C. ternatea flowers. Various phyto-
chemicals such as kaempferol, quercetin and myricetin
glycosides as well as anthocyanins have been isolated from
C. ternatea flowers. Clitoria ternatea flower extracts were
found to possess antimicrobial, antioxidant, anti-inflam-
matory, cytotoxic and antidiabetic activities which are
beneficial to human health. Clitoria ternatea flower is a
promising candidate for functional food applications owing
to its wide range of pharmacotherapeutic properties as well
as its safety and effectiveness.
Keywords Anthocyanin · Antibacterial · Anticancer ·
Anti-inflammatory · Antioxidant · Flavonol
Introduction
Aromatic and medicinal plants have been used for thera-
peutic, religious, cosmetic, nutritional, and beautification
purposes since ancient times and humanity of all civiliza-
tions and culture are familiar with their usage (Senica et al.
2019; Senkal et al. 2019; Gecer et al. 2020). Clitoria ter-natea plant is classified in the kingdom Plantae, phylum
Tracheophyta, class of Magnoliopsida and a family of
Fabaceae (Jamil et al. 2018). Clitoria ternatea is a peren-
nial climber (2–3 m in height) and is known by its common
name as butterfly pea or blue pea flower (Mukherjee et al.
2008). In other regions it is known as aparajita (Bengali),
kajroti (India), cunha (Brazilian), cunha, fula criqua (Por-
tuguese), lan hu die (Chinese), bunga biru, tembang telang
(Indonesian), bunga telang (Malaysian), clitoria azul
(Spanish), dangchan (Thai), chi da˙ˆu biec (Vietnamese) and
mavi kelebek sarmasıgı (Turkish) (Kosai et al. 2015;
Subramanian and Prathyusha 2011; Mukherjee et al. 2008).
It is commonly grown as an ornamental plant and is also
used as a revegetation species while in Southeast Asia, the
blue flower pigment is traditionally used as food colorant
(Havananda and Luengwilai 2019; Oguis et al. 2019) The
plant is known to be suitable as a cover crop and green
manure having the ability to not only suppress perennial
weeds but able to enrich the soil by nitrogen fixation
(Chauhan et al. 2012; Reid and Sinclair 1980). C. ternateaplant is widely distributed in India, Phillipines, in other
tropical Asian countries, South and Central America, the
Caribbean and Madagascar (Sivaranjan and Balachandran
1994; Ambasta 1988).
Clitoria ternatea is considered as a nootropic herb in
Ayurvedic medicine (Chauhan et al. 2017). It grows well in
full sunlight/partially shaded area in which seed germina-
tion takes about 1–2 weeks while 4 weeks is required for
& Wee Sim Choo
[email protected]
1 School of Science, Monash University Malaysia, Jalan
Lagoon Selatan, 47500 Bandar Sunway, Selangor, Malaysia
123
J Food Sci Technol (June 2021) 58(6):2054–2067
https://doi.org/10.1007/s13197-020-04745-3
Page 2
flowering to occur (Jamil et al. 2018; Nguyen et al. 2011).
There are several lines of C. ternatea with different flower
colours of light blue, dark blue, white and mauve which are
4–5 cm long (Fig. 1). Compounds reported to be found in
the flowers are ternatin anthocyanins and various flavanol
glycosides of kaempferol, quercetin and myricetin
(Mukherjee et al. 2008; Kazuma et al. 2003). The leaves
are pinnate with 5–7 leaflets, eliptic-oblong with a length
and width range of 2.5–5.0 and 2.0–3.2 cm. They have flat
linear beaked seed pods with a length range of 5–7 cm and
is edible when tendered. The seed is oval in shape and has a
blackish or yellowish brown colour with a length range of
4.5–7.0 mm and 3–4 mm wide. It has a taproot system with
many slender lateral roots (Kosai et al. 2015; Mukherjee
et al. 2008).
Nutritional analysis of C. ternatea flowers identified the
percentage of protein, fibre, carbohydrate and fat to be
0.32, 2.1, 2.2 and 2.5% respectively while the moisture
content was found to be 92.4%. The flower was also found
to have high content of calcium (3.09 mg/g), magnesium
(2.23 mg/g), potassium (1.25 mg/g), zinc (0.59 mg/g),
sodium (0.14 mg/g) and iron (0.14 mg/g) (Neda et al.
2013). Several studies investigated, identified and isolated
the bioactive compounds from C. ternatea flower. The
anthocyanins known as ternatins are blue in colour and are
acylated based on delphinidin (Fig. 2). Their structures
were characterised as malonylated delphinidin 3,3′,5′-triglucosides having 3′,5′-side chains with alternative D-
glucose and p-coumaric acid units at R and R1 with a total
of 15 (poly) acylated delphinidin glucosides identified in
all the blue petal lines namely ternatins A1-A3, B1-B4, C1-
C4 and D1-D3 while some studies have identified several
other delphinidin derivatives (Zakaria et al. 2018; Shen
et al. 2016; Nair et al. 2015). Ternatins A1, A2, B1, B2, D1
and D2 (Fig. 3) are the six major anthocyanins present in
the flowers (Mukherjee et al. 2008; Terahara et al. 1998).
The flavonols (Fig. 4) identified in the petals are fourteen
kaempferol, quercetin and myricetin glycosides which
consist of H or OH at R1 and R2 and with H, rhamnosyl or
malonyl at R3 and R4 (Mukherjee et al. 2008; Kazuma et al.
2003). Shen et al. (2016) identified various lipophilic
compounds from C. ternatea being fatty acids (palmitic
acid, stearic acids, petroselinic acids, linoleic acid, ara-
chidic acid, behenic acid and phytanic acid), phytosterols
(campesterol, stigmasterol, β-sitosterol and sitostanol) and
tocols (α-tocopherol and -tocopherol). Several other com-
ponents such as mome inositol, pentanal, cyclohexen,
1-methyl-4-(1-methylethylideme) and hirsutene were
identified by Neda et al. (2013). In addition to the identi-
fication of various anthocyanins and flavonol glycosides,
other components such as 6″-malonylastragalin, pheny-
lalanine, coumaroyl sucrose, tryptophan and coumaroyl
glucose were determined (Zakaria et al. 2018).
Clitoria ternatea has been used traditionally in ayurve-
dic medicine for various health issues. Its roots are used to
treat indigestion, constipation, fever, arthritis, sore throat,
skin diseases and eye ailments while its seeds are used as
laxative, to treat colic and swollen joints. The traditional
Cuban culture uses the decoction of the roots alone or
combined with flowers to promote menstruation, induce
uterine contractions as well as to treat liver and intestinal
problems (Mukherjee et al. 2008; Fantz 1991). Various
research studies have been done on the roots, seeds, flowers
and leaves of C. ternatea. Clitoria ternatea flower is knownfor its potential health benefits in which several studies
have shown the crude extract to have antidiabetic (Borikar
et al. 2018), antioxidant (Chayaratanasin et al. 2015),
antimicrobial (Leong et al. 2017) and antiproliferative/an-
ticancer activities (Shen et al. 2016). Thus, C. ternateaflowers can be used as a natural source of antioxidants and/
or a possible supplement in food or pharmaceutical
industries. This paper reviews the current update on the
extraction methods of C. ternatea flowers and its effect on
the phytochemicals as well as the biological activities of
these phytochemicals.
Extraction of phytochemicals
Extraction procedure of phytochemicals from plant mate-
rials is an important step. Various extraction procedures are
available and the identification/selection of optimumFig. 1 Clitoria ternatea flower
Fig. 2 Delphinidin 3-malonyl glucoside
J Food Sci Technol (June 2021) 58(6):2054–2067 2055
123
Page 3
parameters are important to ensure the enhancement of
phytochemical yield (Azmir et al. 2013). Conventional and
non-conventional extraction methods are available having
respective advantages over each other thus careful selec-
tion of method should be evaluated depending on the
suitability of samples and goals needed to be achieved.
Prior to extraction, plant materials are usually reduced in
size to increase the surface area for mixing with solvent
and the samples used are either fresh, dried, grounded or
powdered. Most studies on C. ternatea flowers utilized air/
oven-dried, fresh flowers (Srichaikul 2018; Phrueksanan
et al. 2014; Kamkaen and Wilkinson 2009) or grounded/
powdered, dried flowers (Lakshan et al. 2019; Lopez Prado
et al. 2019; Mehmood et al. 2019; Pham et al. 2019;
Fig. 3 Major anthocyanins in C. ternatea flowers a Ternatin A1, b Ternatin A2, c Ternatin B1, d Ternatin B2, e Ternatin D1, and f Ternatin D2
2056 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 4
Adhikary et al. 2018; Rabeta and An Nabil 2013). Some
studies utilized fresh flowers that were cut into smaller
pieces, washed and stored in −25 °C freezer and extracted
within a month’s time (Chong and Gwee 2015) or freeze
dried followed by grounding (Shen et al. 2016). Several
conventional and non-conventional extraction methods
have been used to obtain phytochemicals from C. ternateaflowers as discussed below.
Conventional extraction
Conventional extraction methods usually involve the use of
different solvents with heat and/or mixing such as soxhlet
extraction, maceration and hydrodistillation which though
effective can be costly and require long extraction time
(Wen et al. 2018; Azmir et al. 2013). Conventional
extraction method is a classical method which has been
widely used for the extraction of C. ternatea flower since
the 1970s. Extraction studies on C. ternatea flower utilising
aqueous solvent mixtures isolated and identified the
structure of various phytochemicals mainly anthocyanins
(Terahara et al. 1989, 1996, 1998) while other studies
(Kazuma et al. 2003; Saito et al. 1985; Ranaganayaki and
Singh 1979) focused on the flavonol constituents. Most
studies employed extractions using aqueous solvent mix-
tures of ethanol or methanol rather than water alone with
heating to investigate its potential bioactivities and phy-
tochemical content while a number of studies investigated
on the optimal solvent and/or extraction parameters
(Table 1).
Ludin et al. (2018) determined the extraction efficiency
of anthocyanins using various solvents (high to low
polarity) in which the ethanol extract was found to have the
highest efficiency for anthocyanin extraction while ethyl
ether extract had the lowest efficiency. The study found
that solvent with higher polarity (ethanol) will have higher
efficiency in extracting polar compounds such as
anthocyanins.
Another study, compared and identified the differences
in the phytochemical content of the flowers using hydro-
philic (methanol) and hydrophobic (ethyl acetate and
hexane; 1:1) extraction. The hydrophilic extract contained
various anthocyanins, kaempferol and quercetin glycosides
while the hydrophobic extract was composed of fatty acids,
phytosterols and tocopherols showing the effect of com-
pounds extracted depending on polarity of compounds
(Shen et al. 2016).
Studies investigating the effect of pH for anthocyanin
extraction found lower pH to be optimum (Mauludifia et al.
2019; Ludin et al. 2018) suggesting lower pH enhances the
extraction of anthocyanins as increase in acidity changes
anthocyanins to the flavium cation form (more stable)
which may enhance vacuole cell wall breakage, increase
solubility of pigments thus increasing the concentration of
anthocyanins extracted. However, the possibility of inhi-
bition of enzymatic oxidation of phenolics as well as the
maintenance of the stability of extracted anthocyanins at
low pH (Zhang et al. 2001) was suggested as a possible
factor as well (Ruenroengklin et al. 2008).
Investigation of the extraction temperature for antho-
cyanin extraction was found to increase with an increase in
temperature and 70 °C was found to be the optimum
temperature (Ludin et al. 2018). The increase in extraction
temperature facilitates higher extraction of anthocyanins as
it increases the internal energy of the molecules which
increases the diffusion and solubility of the pigments thus
having a higher yield (Cacace and Mazza 2003). However,
there was reduction in anthocyanin concentration at 80 °Cwhich may be due to degradation of pigments (Ludin et al.
2018). A particular study compared the effect of extraction
with water or aqueous methanol which found the latter to
have a higher phenolic content. However, both extractions
differed in time and temperature which may have influ-
enced the outcome of the results thus suitable controls are
required to have a fair justification and comparison of
extraction variables (Rabeta and An Nabil 2013).
Several studies utilised the Response Surface Method-
ology (RSM) which adapts a multivariate system that fits
experimental data in a statistical model by a response
function to optimise the extraction of phytochemical of
interest with variation of experiment parameters (temper-
ature, extraction time, liquid–solid ratio). The utilisation of
RSM enabled the identification of optimum parameters for
the extraction of anthocyanins (114–132 mg/L), extract
yield (23.7–53.4%) and phenolic content (594–692 mg
GAE/100 g and 24–81 mg GAE/L) (Escher et al. 2020;
Pham et al. 2019; Baskaran et al. 2019; Lakshan et al.
2019). Studies utilising RSM to enhance the extraction of
phytochemical of interest is beneficial as it incorporates
variables of interest (e.g. temperature, extraction time,
liquid-solvent ratio) to be studied using a statistical
approach which generates a mathematical model that
optimizes extraction variables (Sreejith et al. 2014).
Fig. 4 Flavonol glycosides
J Food Sci Technol (June 2021) 58(6):2054–2067 2057
123
Page 5
Table 1 The extraction of phytochemicals from C. ternatea flowers
Extraction solvent Phytochemical extracted Optimal extraction
condition
References
Ethanol, methanol, chloroform, acetonitrile,
acetone, ethyl acetate, n-butyl, water,
n-hexane and ethyl ether
Anthocyanins Ethanol Ludin et al.
(2018)
70% ethanol
(40–80 °C)
70 °C
70% ethanol
(pH 4–9)
pH 4
Water
(pH 2, 7 and 10)
Anthocyanins pH 2 Mauludifia
et al.
(2019)
Methanol Anthocyanins (Ternatin A1, B2, B3, C2, D2, D3,
delphinidin derivatives), kaempferol
3-neohesperidoside and quercetin 3-(2G-
rhamnosylrutinoside), rutin, ellagic acid
– Shen et al.
(2016)
Ethyl acetate and hexane (1:1) Fatty acids (palmitic acid, stearic acids, petroselinic
acids, linoleic acid, arachidic acid, behenic acid and
phytanic acid), phytosterols (campesterol,
stigmasterol, β-sitosterol and sitostanol) and tocols
(α-tocopherol and -tocopherol)
–
50% ethanol
(40–80 °C, 15–75 min, liquid–solid ratio of
10:1 to 30:1 mL/g)
Anthocyanins 60.6 °C, 6 min, liquid–
solid ratio of 23:1 mg/
L
Pham et al.
(2019)
Water
(25–95 °C, 40–80 min liquid–solid ratio of
20:1 to 60:1)
Extract yield 54 °C, 74 min, liquid–
solid ratio of 37:1
Baskaran
et al.
(2019)
Water
(40–80 °C, 30–60 min liquid–solid ratio of 1
to 3 g/L)
Phenolics 59.6 °C, 37 min, liquid–
solid ratio of 3 g/L
Lakshan
et al.
(2019)
Water
(11.7–68.3 °C, 8.78–51.21 min)
Phenolics 40 °C, 30 min Escher
et al.
(2020)
40% and 50% ethanol
(with 30 min ultrasound or maceration alone
for 1–7 days)
Phenolics 50% ethanol with
30 min ultrasound
Srichaikul
(2018)
Flavonoids Maceration in 40%
ethanol for 7 days
Water
(30–50 °C, 30–150 min, 96–240 W, liquid–
solid ratio of 2–15 mL/g)
Anthocyanins 50 °C, 150 min, liquid–
solid ratio of 15 mL/g
and 240 W
Chong and
Gwee
(2015)
Water with ultrasound
(50 °C, 150 min, liquid–solid ratio of 15 mL/
g and 240 W)
95% ethanol without ultrasound
(50 °C, 150 min, liquid–solid ratio of 15 mL/
g)
Water with ultrasound
assistance
Water with and without ultrasound
(50 °C, 150 min, liquid–solid ratio of 15 mL/
g and 240 W)
Phenolics and flavonoids Water with ultrasound
assistance
Mehmood
et al.
(2019)
Water with heating (3 h)
Water with microwave (2 min)
Dye (anthocyanins) Water with microwave
assistance
Sinha et al.
(2012)
2058 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 6
Non-conventional extraction methods
Non-conventional extraction methods are newer, highly
efficient, safer to the environment having various advan-
tages over conventional extraction method which include
methods such ultrasound assisted extraction, microwave
assisted extraction, enzyme assisted extraction, supercriti-
cal fluid extraction and pressurized liquid extraction (Wen
et al. 2018; Azmir et al. 2013). To the author’s best
knowledge, among the non-conventional extraction meth-
ods, only ultrasound and microwave assisted extraction
have been employed to extract phytochemicals from C.ternatea flowers (Table 1).
Ultrasound assisted extraction works on the concept of
acoustic waves production leading to molecular movement
of solvent and sample which facilitates the leaching of
organic and inorganic compounds (Herrera and De Castro
2005). Srichaikul (2018) compared the effect of short
extraction time with ultrasound and long extraction time
with maceration for 1–7 days for extraction efficiency of
phenolic and flavonoid content using aqueous ethanol.
Extraction of phenolic content was more efficient in the
ultrasound assisted extraction. However, for flavonoid
content, maceration in aqueous ethanol at day 7 was found
to be higher than with ultrasound extraction (30 min).
Although the flavonoid content was higher at day 7 in the
aqueous ethanol extract without ultrasound assistance, a
longer extraction time (7 days) was required to achieve this
effect. A particular study utilised RSM to obtain the opti-
mum extraction condition for anthocyanins using water by
varying the extraction time, temperature, liquid–solid ratio
and ultrasound frequency. The extraction using the opti-
mized condition showed higher extraction yield of antho-
cyanins by 246.5% than 95% methanol without ultrasound
assistance (Chong and Gwee 2015). Mehmood et al. (2019)
further investigated this optimized condition by Chong and
Gwee (2015) to determine the extraction efficiency of
phenolic and flavonoid content from C. ternatea flowers by
using water with and without ultrasound assistance. Simi-
larly, ultrasound extraction showed a better extraction yield
of phenolic and flavonoid content.
In microwave assisted extraction, electromagnetic
energy is converted to heat following ionic conduction and
dipole rotation mechanisms which promotes the release of
solutes from the sample matrix to solvent (Alupului et al.
2012; Jain 2009). Sinha et al. (2012) compared the
extraction efficiency of C. ternatea dye (anthocyanins)
using conventional extraction with water and heating to
microwave assisted extraction. Microwave assisted
extraction was more efficient than conventional extraction
having a higher dye yield. Microwave assisted extraction
was also more time efficient requiring 2 min as opposed to
conventional extraction method which required 3 h.
Although conventional extraction has widely been used
for the extraction of these flowers, the use of non-con-
ventional extraction method (ultrasound assistance) has
shown to be superior and beneficial for the extraction of
phytochemicals. Thus exploration on the use of other non-
conventional extraction methods which are considered as
“green techniques” would be beneficial in determining
extraction efficiency of various phytochemicals. Other
studies have employed the use of ultrasound, pulsed-elec-
tric field, pressurized liquid and microwave assisted
extraction which were more effective than the conventional
extraction method for extraction of phenolics and antho-
cyanins which required shorter extraction time and were
also useful in preventing oxidation of compounds (Caldas
et al. 2018; Corrales et al. 2008, 2009).
Biological activities
Clitoria ternatea flower contains a significant amount of
phytochemicals which exhibits great antioxidant, antimi-
crobial, antidiabetic, anti-inflammatory and antiprolifera-
tive/anticancer properties (Lopez Prado et al. 2019;
Mahmad et al. 2018; Nair et al. 2015; Rajamanickam et al.
2015; Neda et al. 2013). Acute toxicity study using albino
Wistar rats treated orally with aqueous ethanol extract
(2000 mg/kg bodyweight) of the flower showed no signs of
mortality or abnormality and there was no significant dif-
ference in the haematological values The extract did not
display acute toxicity effects and are safe for consumption
(Srichaikul 2018). Clitoria ternatea flowers can potentially
be utilised as a functional food incorporated into various
food products or even as a pharmaceutical supplement/drug
combined with commercial drugs to improve treatment
efficacy of patients.
Antioxidant activity
Oxidative stress plays a part in the development of chronic
and degenerative illness such as cancer, autoimmune dis-
orders, cardiovascular and neurodegenerative diseases. The
discovery of antioxidants from natural sources is beneficial
to human health (Admassu et al. 2018; Pham-Huy et al.
2008). Various studies investigated the antioxidant activity
of C. ternatea flowers using antioxidant assays such as 2,2-
diphenyl-1-picrylhydrazyl radical (DPPH) radical scav-
enging, ferric reducing antioxidant power (FRAP), hydro-
xyl radical scavenging activity (HRSA), hydrogen peroxide
scavenging, oxygen radical absorbance capacity (ORAC),
superoxide radical scavenging activity (SRSA), ferrous ion
J Food Sci Technol (June 2021) 58(6):2054–2067 2059
123
Page 7
chelating power, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging and Cu2+
reducing power assays. Clitoria ternatea flower has been
shown to have potent antioxidant activity (Table 2).
In the DPPH assay, 100% methanol extract of C. ter-natea flower extract was found to be more potent than
vitamin E (Nithianantham et al. 2013), whereas the water
extract was found to be lower than ascorbic acid (vitamin
C) (Chayaratanasin et al. 2015; Phrueksanan et al. 2014;
Iamsaard et al. 2014) Some studies investigated and com-
pared the antioxidant activity (DPPH assay) of the extracts
using different solvents in which the water extract was
found to be more potent than 100% ethanol extract at
15 min extraction time (Kamkaen and Wilkinson 2009).
However, in another study the best extraction time was
determined (6 h) for the water extract, 100% and 50%
methanol extract in which the water extract and 50%
methanol were found to be equally potent and had a higher
activity than 100% methanol extract (Lopez Prado et al.
2019). The optimum condition was investigated using
water extract in another study with and without ultrasound
at a fixed temperature and liquid-solvent ratio in which the
extraction with ultrasound was found to have higher
antioxidant activity (Mehmood et al. 2019). The in vitro
chemical assays to measure antioxidant activity (Table 2)
need to be carefully interpreted as they bear no similarity to
biological systems including the absorption of antioxidants
by the human body (Gengatharan et al. 2015). In a cell
based study, the water extract was found to potently inhibit
2,2′-azobis-2-methyl-propanimidamide dihydrochloride
(AAPH)-induced hemolysis and oxidative damage of
canine erythrocytes (Phrueksanan et al. 2014). In another
study, the pre-treatment of human HaCaT keratinocytes
with the water extract reduced UV-induced mitochondrial
DNA damage (Zakaria et al. 2018). In a randomised
crossover study, acute ingestion of C. ternatea flower
extract/beverage was found to have increased plasma
antioxidant capacity and the effect was further enhanced
when consumed together with sucrose in healthy men
(Chusak et al. 2018). These studies attribute the flavonols
and anthocyanins for the antioxidant activity.
Antibacterial activity
The emergence of antibiotic resistance microbes limits the
effectiveness of current drugs significantly causing treat-
ment failure of infections (Scheffler et al. 2013). In regard
to this challenge, there is a need to develop alternative
approaches in addition to searching for new antibacterial
compounds. In vitro activity of an antimicrobial (antibac-
terial or antifungal) agent can be tested through various
methods such as broth or agar dilution, and disc diffusion
methods (Balouiri et al. 2016). Several studies investigated
on the antibacterial potential of C. ternatea flowers. The
methanol extract of C. ternatea flower was tested against
12 bacterial species (Bacillus cereus, Bacillus subtilis,Bacillus thuringiensis, Staphylococcus aureus, Streptococ-cus faecalis, Escherichia coli, Klebsiella pneumoniae,Pseudomonas aeruginosa, Salmonella typhi, Enterobacteraerogens, Proteus mirabilis and Herbaspirillum spp.) and
was found to have the most potent activity against Bacillusthuringiensis with a minimum inhibitory concentration
(MIC) of 12.5 mg/mL and minimum bactericidal concen-
tration (MBC) of 25 mg/mL with an inhibition zone of
15.7 mm using agar disc diffusion technique (Kamilla et al.
2009). In another study, the water, methanol, petroleum
ether, hexane and chloroform extract of C. ternatea flower
(4 mg) were tested against E. coli, K. pneumoniae, S.enteritidis, S. typhimurium and P. aeruginosa to determine
its antibacterial activity. The methanol extract was found to
have the highest activity when tested using agar disc dif-
fusion technique with an inhibition zone range of 16–
26 mm in E. coli, K. pneumonia and P. aeruginosa but had
no activity against S. typhi and S. enteritidis. The highest
zone of inhibition 26 mm was observed against K. pneu-monia and P. aeruginosa (Uma et al. 2009). Leong et al.
(2017) determined the antibacterial activity of antho-
cyanins of C. ternatea flower ethanol extract paste against
B. cereus, B. subtilis, S. aureus, B. subtilis subsp. spizizenii,Proteus mirabilis, K. pneumoniae, Yersinia enterocoliticaand E. coli. The extract was found to have good antibac-
terial activity against B. cereus, B. subtilis, S. aureus, P.mirabilis and K. pneumonia with the most potent activity
against K. pneumonia with a MIC of 1.6 mg/mL and
minimum lethal concentration (MLC) of 25 mg/mL while
in another study, the anthocyanin fraction obtained from
the ethanol extract of C. ternatea flower had the best effect
against B. subtilis with a disc diffusion inhibition zone of
10 mm (Mahmad et al. 2018). The findings from these
studies suggest the potential of the anthocyanins for its
antibacterial activity.
Antifungal activity
A rise in resistance towards most antifungal agents in
diverse pathogens which calls for the need to identify new
therapeutic agents (Perfect 2016). The methanol extract of
C. ternatea flower (100 mg/mL) tested against Candidaalbicans, Rhizopus and Penicillium spp. had the highest
activity against Candida albicans with an inhibition zone
of 19 mm in agar disc diffusion. However, in broth dilution
method, it only had activity against Penicillium spp. and
Rhizopus with similar MIC value of 0.8 mg/mL and MFC
value of 1.6 mg/ml (Kamilla et al. 2009). The anthocyanin
2060 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 8
Table 2 The antioxidant activity of C. ternatea flowers from various research studies
Extract Antioxidant assay Results References
Water extract and 100% ethanol extract DPPH radical scavenging Water extract IC50=1 mg/mL
Ethanol extract IC50=4 mg/mL
Water extract in gel formulation for
inhibition of DPPH reduction=28% at
0.5 mg/mL
Kamkaen and
Wilkinson
(2009)
100% Methanol extract DPPH radical scavenging IC50=327 µg/mL Nithianantham
et al. (2013)
Citrate buffer extract DPPH radical scavenging EC50=0.49 mg/mL Siti Azima et al.
(2014)
FRAP 13.3 mM/g based on trolox equivalent
antioxidant capacity (TEAC)
Water extract DPPH radical scavenging IC50=84.15 µg/mL Iamsaard et al.
(2014)
FRAP 0.33 mmol/mg ascorbic equivalent
Water extract DPPH radical scavenging IC50=470 µg/mL Phrueksanan
et al. (2014)
Oxygen radical absorbance
capacity (ORAC)
17.54 g trolox equivalents/mg extract
Reduction of free radical-
induced erythrocyte
hemolysis (4 h)
96.3% at 400 µg/mL
Inhibition of lipid
peroxidation (4 h)
72.7% at 400 µg/mL
Water extract DPPH radical scavenging IC50=0.47 mg/mL Chayaratanasin
et al. (2015)
Trolox equivalent antioxidant
capacity (TEAC)
0.17 mg trolox/mg dried extract
Ferric reducing antioxidant
power (FRAP)
0.38 mmol FeSO4/mg dried extract
HRSA IC50=19.2 mg/mL
SRSA IC50=26.3 mg/mL
Ferrous ion chelating power [103 mg EDTA/mg dried extract
Methanol/acetone/water
(5:4:1) extract
ORAC 490.7 mol trolox equivalent/g extract Nair et al.
(2015)
95% methanol extract DPPH radical scavenging IC50=95.3 µg/mL Rajamanickam
et al. (2015)
Water extract ABTS radical scavenging 4.2 µM trolox equivalent/g extract Azima et al.
(2017)
DPPH radical scavenging EC50=0.76 mg/mL
FRAP 10.9 mM trolox equivalent/g extract
ORAC 15.8 µmol trolox equivalent/g extract
Water extract ABTS radical scavenging IC50=42.9 µg/mL Zakaria et al.
(2018)
DPPH radical scavenging IC50=195.5 µg/mL
Water extract with ultrasound assistance (US)
and water extract with heat assistance at 50 °C(AGE)
DPPH radical scavenging US=931.5 µg trolox equivalent/g extract
AGE=764.3 µg trolox equivalent/g
extract
Mehmood et al.
(2019)
ABTS radical scavenging US=13,488 µg trolox equivalent/g extract
AGE=11,720.3 µg trolox equivalent/g
extract
FRAP US=5834.6 µg trolox equivalent/g extract
AGE=4195.3 µg trolox equivalent/g
extract
J Food Sci Technol (June 2021) 58(6):2054–2067 2061
123
Page 9
fraction obtained from the ethanol extract of C. ternateaflower tested against Fusarium sp., A. niger and Tricho-derma sp. had the highest activity against Fusarium sp.
with an inhibition zone of 10 mm in agar disc diffusion
technique (Mahmad et al. 2018). The anthocyanins of C.ternatea flower ethanol extract paste (50 mg/mL) tested
against A. niger, P. expansum and R. stolonifera only
exhibited activity against P. expansum with an inhibition
zone of 15.5 mm in agar disc diffusion while it had an MIC
value of 12.5 mg/mL and MLC value of 25 mg/mL. The
mode of action for the antifungal activity against P.expansum was investigated and found to be mediated by
the alteration of morphology of P. expansum fungal hyphae
which had flattened empty hyphae resulting from cell wall
disruption and damage of conidiophore. The germination
of P. expansum conidia was completely inhibited with
suppressed conidial development (Leong et al. 2017).
Anti-inflammatory activity
The current available non-steroidal anti-inflammatory
drugs (NSAIDs), including acetaminophen and aspirin are
associated with side effects, particularly gastrointestinal
and cardiovascular effects as they are known to affect both
COX-1 and COX-2. The discovery of new or alternate
strategies is needed to reduce the risks associated with
NSAIDs while achieving sufficient pain relief (Brune and
Patrignani 2015).
The petroleum ether extract of C. ternatea flowers was
evaluated for anti-inflammatory activity using carrageenan
paw edema method with healthy albino rats of either sex.
The extract (200 and 400 mg/kg) significantly inhibited
paw edema compared to control untreated group while in
Eddy’s hot plate method, the treatment group (400 mg/kg)
had significant increased reaction time (time recorded
when animals licked their fore or hind paws or jump
response, whichever appear first) compared to control
untreated group. The study suggests the possibility of the
extracts to have a protective effect against the release of
prostaglandins, kinnins and other substances in car-
rageenan induced edema (Shyamkumar and Ishwar 2012).
In another study, the anthocyanin and flavonol fraction
obtained from C. ternatea flower extract (extracted in a
mixture of MeOH/acetone/H2O at ratio of 5:4:1) were
investigated for its anti-inflammatory potential. In the
lipopolysaccharides (LPS)-induced inflammation in RAW-
264.7 macrophage cells, the flavonols had mild suppression
of ROS while the anthocyanins had no effect on ROS
production. The anthocyanins were also found to have
higher inhibition of nitric oxide production compared to the
flavonols. In western blot studies, only the anthocyanins
inhibited nuclear factor-B translocation and iNOS protein
expression whereas the flavonols significantly inhibited
COX-2 expression but not the anthocyanins (Nair et al.
2015).
Table 2 continued
Extract Antioxidant assay Results References
Reducing power US=4539.0 µg trolox equivalent/g extract
AGE=6154.1 µg trolox equivalent/g
extract
Cu2+ reducing power US=12,696 µg trolox equivalent/g extract
AGE=9549 µg trolox equivalent/g extract
Xanthine oxidase inhibition US=1.01 mg/mL (IC50)
AGE=1.22 mg/mL (IC50)
Water extract, 100% and 50% methanol extract
at 6 h (best condition)
DPPH radical scavenging Water extract=11.7 mM trolox
equivalent/g extract
100% methanol extract=6.99 mM trolox
equivalent/g extract
50% methanol extract=12.2 mM trolox
equivalent/g extract
Lopez Prado
et al. (2019)
Inhibition of cholesterol
oxidation
Water extract=79.8%
100% methanol extract=49.7%
50% methanol extract=89.8%
2062 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 10
Cytotoxic and anti-proliferative/anticanceractivities
Chemotherapy, radiation therapy and targeted therapy are
among the approaches used for the treatment and man-
agement of cancer but they are not able to provide a per-
manent cure and have been associated with various side
effects and toxicities (Curigliano et al. 2012). Thus, new
agents that are safe, available and effective are urgently
needed. Several studies investigated the anticancer poten-
tial of C. ternatea flower extracted using different solvents.
The 100% petroleum ether extract (IC50=36 µg/mL) was
found to be more potent than the 100% ethanol extract
(IC50 value of 57 µg/mL) in the in vitro cytotoxic assay
against Dalton’s lymphoma ascites (DLA) cells at 3 h
which could be due to different phytochemical composition
in both extracts. The petroleum ether extract was found to
have presence of saponins, tannins, steroids and triterper-
noids while the ethanol extract had flavonols only (Kumar
and Bhat 2011). In another study, the water extract was
more potent than the methanol extract having much lower
IC50 values with activity against hormone dependent breast
cancer cell line (MCF-7), non-hormone-dependent breast
cancer cell line (MDA-MB-231), human ovary cancer cell
line (Caov-3), and human liver cancer cell line (HepG2) at
72 h. However, the methanol extract had activity against
MCF-7 and MDA-MB-231 cells only with higher IC50
values. The extracts were not toxic against the normal cell
line (Hs27). The study suggests the aqueous extract to have
more significant anti-proliferative activity than the metha-
nol extract as it may have more active compounds (flavo-
noids) present (Neda et al. 2013). Shen et al. (2016) found
the anticancer effect of the hydrophilic (100% methanol)
extract to be more potent than the lipophilic (hexane:ethyl
acetate, 1:1) extract on human epithelial laryngeal carci-
noma (Hep-2) cell line. The potent active compounds
identified in the hydrophilic extract were mainly ternatins,
kaempferol and quercetin responsible for the antiprolifer-
ative effect as opposed to the lipophilic extract which
constitutes of fatty acids, phytosterols and tocopherols.
Antidiabetic activity
Oral antidiabetic medications such as biguanides, megli-
tinide, thiazolidinedione, sulfonylureas and dipeptidyl
peptidase 4 are known to be associated with various side
effects (Chaudhury et al. 2017). Herbal based medications
are worth exploring for potential use in the management of
diabetes as they are considered to be safer and may have
reduced side effects (Borikar et al. 2018). Several studies
investigated the in vitro and in vivo potential of C. ternatea
flower extract for antidiabetic activity. The water extract
reduced the formation of fluorescent advanced glycation
end products having the highest activity at day 28 (49.4%
at 1 mg/mL) as well as significant reduction in fruc-
tosamine level (14.47–36.66%) in glycated bovine serum
albumin. The study suggests the potential of the extract in
the prevention of the formation of advanced glycation end
products to be mediated through its free radical scavenging
ability mainly attributed to the active compounds present
being the ternatin anthocyanins, delphinidin derivatives
and kaempferol (Chayaratanasin et al. 2015). In vivo study
for antidiabetic activity in alloxan-induced diabetic rats
(wistar albino) by Rajamanickam et al. (2015) utilising
95% methanol, ethyl acetate and chloroform extract were
found to have significantly reduced blood glucose level,
increased serum protein levels and restored serum albumin
to normal levels. The extracts also significantly decreased
serum urea, creatinine, cholesterol and triglyceride levels
compared to control untreated diabetic rats. A similar trend
was also observed in the in vivo studies by Borikar et al.
(2018) utilising 100% methanol extract and water extract in
the study by Daisy and Rajathi (2009). In a randomised
crossover study, acute ingestion of C. ternatea flower
extract/beverage was found to have suppressed postpran-
dial plasma glucose and insulin levels when consumed with
sucrose in healthy men (Chusak et al. 2018). Overall, these
studies suggested the hypoglycemic activity may be exer-
ted by the flavonoid principles (flavonol glycosides and
anthocyanins) and alkaloids present in the extract which
may involve the potentiation of insulin secretion from the
β-cell or by enhancement of the transport of blood glucose
from plasma to peripheral tissues.
Other biological activities of phytochemicalsstudied in C. ternatea flowers
Clitoria ternatea flower has shown to have potential
antioxidant, antimicrobial, anticancer and antidiabetic
activity. However, there are also various studies which
have looked into its potential for other beneficial activities.
Adhikary et al. (2018) found the 100% methanol extract of
C. ternatea flower and its purified compound quercetin-3β-D-glucoside for its anti-arthritic potential in a mice model.
Quercetin-3β-D-glucoside was found to be more potent
than the extract to significantly reduce myeloperoxidase
activity, decrease in release of pro-inflammatory cytokines,
chemokines, reactive oxygen species (ROS)/reactive
nitrogen species production. It also significantly reduced
tumor necrosis factor α-receptor 1, toll-like receptor 2,
inducible isoform of nitric oxide synthase, COX-2 and
matrix metalloproteinase-2 expression.
J Food Sci Technol (June 2021) 58(6):2054–2067 2063
123
Page 11
The anti-allergy effects of C. ternatea flower extract wasalso found in a study by Singh et al. (2018). The 98%
ethanol extract was able to attenuate histamine-induced
contraction in both goat tracheal chain and isolated guinea
pig ileum preparations. The extract was also found to
attenuate histamine-induced dyspnoea and ovalbumin-in-
duced changes of various inflammatory cytokines in animal
models. The extract also displayed antitussive activity in
sulfur dioxide- and citric acid-induced cough in experi-
mental animals and attenuated inflammation in carrageenan
and acetic acid challenged rodents. Clitoria ternatea flower
extract was also found to have other beneficial effects in
various other studies such as anthelmintic (Nirmal et al.
2008), larvicidal (Mathew et al. 2009), anti-aging (Zakaria
et al. 2018), hepatoprotective (Nithianantham et al. 2013),
testicular damage protection (Iamsaard et al. 2014), anti-
adipogenesis (Chayaratanasin et al. 2019) and starch
digestion (Chusak et al. 2019) activity.
Conclusion
Clitoria ternatea is a versatile plant known for its tradi-
tional application in ayurvedic medicine, food colourant
and cover crop among others. Various beneficial studies
have been done on this plant along the years which have
found it to have many health benefits thus giving a greater
insight on its potential uses. Extraction studies have been a
very important step in identifying variables which influ-
ences the extraction of phytochemicals. Conventional
extraction such as the use of solvent for maceration of
samples has been the most common approach and shown to
be efficient. However, several disadvantages have been
associated with this method due to high cost, requiring long
extraction time and is not environmentally friendly with the
use of solvent. Thus newer studies have explored and
compared extraction methods such as ultrasound/mi-
crowave assisted extraction (non-conventional method)
which are not only environmentally friendly but are much
more time efficient than conventional extraction methods.
Thus, future studies could be geared towards the use of
non-conventional extraction for C. ternatea flowers as well
as exploring other environmentally safe methods such as
pulsed-electric field, pressurized liquid, enzyme-assisted
and pressurised liquid extraction. Findings from acute
toxicity studies for C. ternatea flowers are rather encour-
aging as it has shown to be safe for consumption in vivo
studies. Thus it is recommended for further studies to
evaluate it for subchronic/chronic toxicity, histopathology
as well as clinical studies to determine its effect and safety
for long term consumption. Most studies have shown the
phytochemicals in C. ternatea flowers, mainly antho-
cyanins, quercetin and kaempferol glycosides to probably
be responsible for the beneficial biological effects. There
are variants of anthocyanins, quercetin and kaempferol
present in the flowers. Thus it would be beneficial for
future studies to explore and compare the effect between
anthocyanin/flavonol fractions which would help guide the
potential fraction to further isolate the compound most
probably responsible for the studied effect. Cytotoxicity
studies have shown the extract to be more selective towards
cancer cell lines than normal cells which may have less
side effects in the human body. Some studies have deter-
mined the mode of action of these bioactive constituents
which provides a lead in understanding the causative bio-
logical effect observed. Exploring the mechanism of action
of the extract/active compounds in eliciting the biological
effect are crucial to understand how it may affect/modulate
certain pathways/molecular targets in the human body and
are thus warranted in further studies. Numerous studies
have shown C. ternatea flower for its potential antioxidant
activity not only in chemical based assays but in cell based
assays and in vivo studies. The consumption of C. ternateaflower extract/beverage was shown to have potential
antioxidant and antihyperglycemic effects in human sub-
jects. However, the effect was only studied in healthy
subjects which may not be generalised to all population.
Future studies looking on the effect on subjects with par-
ticular health conditions compared to healthy subjects are
recommended to better understand its effect and determine
its potential. Owing to the many benefits displayed by this
flower, it is worthwhile to carry out suggested further
studies to better understand the biological effects which
have been reported so far as well as exploring other
bioactivities. The bioactive compounds of C. ternateaflower are promising candidates for development as novel
and effective pharmaceutical agents as well as applications
as functional foods to promote human health and
wellbeing.
Acknowledgement This work was funded by the School of Science,
Monash University Malaysia.
References
Adhikary R, Sultana S, Bishayi B (2018) Clitoria ternatea flower
petals: effect on TNFR1 neutralization via downregulation of
synovial matrix metalloproteases. J Ethnopharmacol 210:209–
222. https://doi.org/10.1016/j.jep.2017.08.017
Admassu H, Gasmalla MA, Yang R et al (2018) Bioactive peptides
derived from seaweed protein and their health benefits: antihy-
pertensive, antioxidant, and antidiabetic properties. J Food Sci
83:6–16. https://doi.org/10.1111/1750-3841.14011
Alupului A, Calinescu I, Lavric V (2012) Microwave extraction of
active principles from medicinal plants. UPB Sci Bull Ser B
74:129–142
Ambasta SP (1988) The Wealth of India: A Dictionary of India Raw
Materials and Industrial Products, vol. II. Publication and
2064 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 12
Information Directorate, CSIR, New Delhi, India, p. 233. ISBN:
8185038902
Azima AS, Noriham A, Manshoor N (2017) Phenolics, antioxidants
and color properties of aqueous pigmented plant extracts: Ardisiacolorata var. elliptica, Clitoria ternatea, Garcinia mangostanaand Syzygium cumini. J Funct Foods 38:232–241. https://doi.org/10.1016/j.jff.2017.09.018
Azmir J, Zaidul ISM, Rahman MM et al (2013) Techniques for
extraction of bioactive compounds from plant materials: a
review. J Food Eng 117:426–436. https://doi.org/10.1016/j.
jfoodeng.2013.01.014
Balouiri M, Sadiki M, Ibnsouda SK (2016) Methods for in vitro
evaluating antimicrobial activity: a review. J Pharm Anal 6:71–
79. https://doi.org/10.1016/j.jpha.2015.11.005
Baskaran A, Mudalib SKA, Izirwan I (2019) Optimization of aqueous
extraction of blue dye from butterfly pea flower. J Phys Conf Ser
1358:012001. https://doi.org/10.1088/1742-6596/1358/1/012001
Borikar SP, Kallewar NG, Mahapatra DK et al (2018) Dried flower
powder combination of Clitoria ternatea and Punica granatumdemonstrated analogous anti-hyperglycemic potential as com-
pared with standard drug metformin: in vivo study in Sprague
Dawley rats. J Appl Pharm Sci 8:75–79. https://doi.org/10.7324/
japs.2018.81111
Brune K, Patrignani P (2015) New insights into the use of currently
available non-steroidal anti-inflammatory drugs. J Pain Res
8:105. https://doi.org/10.2147/jpr.s75160
Cacace JE, Mazza G (2003) Mass transfer process during extraction
of phenolic compounds from milled berries. J Food Eng 59:379–
389. https://doi.org/10.1016/s0260-8774(02)00497-1
Caldas TW, Mazza KE, Teles AS et al (2018) Phenolic compounds
recovery from grape skin using conventional and non-conven-
tional extraction methods. Ind Crop Prod 111:86–91. https://doi.
org/10.1016/j.indcrop.2017.10.012
Chauhan NS, Singh NK, Gupta JK et al (2017) A Review on Clitoria
ternatea (Linn.): Chemistry and Pharmacology. Medicinal Plants
and its Therapeutic Uses. OMICS Group eBooks, CA, USA.
ISBN: 1632780747
Chaudhury A, Duvoor C, Dendi R et al (2017) Clinical review of
antidiabetic drugs: implications for type 2 diabetes mellitus
management. Front Endocrinol 8:6. https://doi.org/10.3389/
fendo.2017.00006
Chauhan N, Rajvaidhya S, Dubey BK (2012) Pharmacognostical,
phytochemical and pharmacological review on Clitoria ternateafor antiasthmatic activity. Int J Pharm Sci Res 3:398
Chayaratanasin P, Barbieri MA, Suanpairintr N et al (2015) Inhibitory
effect of Clitoria ternatea flower petal extract on fructose-
induced protein glycation and oxidation-dependent damages to
albumin in vitro. BMC Complement Altern Med 15:27. https://
doi.org/10.1186/s12906-015-0546-2
Chayaratanasin P, Caobi A, Suparpprom C et al (2019) Clitoriaternatea flower petal extract inhibits adipogenesis and lipid
accumulation in 3T3-L1 preadipocytes by downregulating
adipogenic gene expression. Molecules 24:1894. https://doi.
org/10.3390/molecules24101894
Chong FC, Gwee XF (2015) Ultrasonic extraction of anthocyanin
from Clitoria ternatea flowers using response surface method-
ology. Nat Prod Res 29:1485–1487. https://doi.org/10.1080/
14786419.2015.1027892
Chusak C, Thilavech T, Henry CJ et al (2018) Acute effect of Clitoriaternatea flower beverage on glycemic response and antioxidant
capacity in healthy subjects: a randomized crossover trial. BMC
Complement Altern Med 18:1–11. https://doi.org/10.1186/
s12906-017-2075-7
Chusak C, Ying JAY, Zhien JL et al (2019) Impact of Clitoriaternatea (butterfly pea) flower on in vitro starch digestibility,
texture and sensory attributes of cooked rice using domestic
cooking methods. Food Chem 295:646–652. https://doi.org/10.
1016/j.foodchem
Corrales M, Toepfl S, Butz P et al (2008) Extraction of anthocyanins
from grape by-products assisted by ultrasonics, high hydrostatic
pressure or pulsed electric fields: a comparison. Innov Food Sci
Emerg Technol 9:85–91. https://doi.org/10.1016/j.ifset.2007.06.
002
Corrales M, Garcıa AF, Butz P et al (2009) Extraction of
anthocyanins from grape skins assisted by high hydrostatic
pressure. J Food Eng 90:415–421. https://doi.org/10.1016/j.
jfoodeng.2008.07.003
Curigliano G, Cardinale D, Suter T et al (2012) Cardiovascular
toxicity induced by chemotherapy, targeted agents and radio-
therapy: ESMO Clinical Practice Guidelines. Ann Oncol 23:55–
66. https://doi.org/10.1093/annonc/mds293
Daisy P, Rajathi M (2009) Hypoglycemic effects of Clitoria ternateaLinn. (Fabaceae) in alloxan-induced diabetes in rats. Trop J
Pharm Res 8:393–398. https://doi.org/10.4314/tjpr.v8i5.48082
Escher GB, Marques MB, do Carmo MAV et al (2020) Clitoriaternatea L. petal bioactive compounds display antioxidant,
antihemolytic and antihypertensive effects, inhibit α-amylase
and α-glucosidase activities and reduce human LDL cholesterol
and DNA induced oxidation. Food Res Int 128:108763. https://
doi.org/10.1016/j.foodres.2019.108763
Fantz PR (1991) Ethnobotany of Clitoria (Leguminosae). Econ Bot
45:511–520. https://doi.org/10.1007/BF02930715
Gecer MK, Kan T, Gundogdu M et al (2020) Physicochemical
characteristics of wild and cultivated apricots (Prunus armeniacaL.) from Aras valley in Turkey. Genet Resour Crop Eviron
67:935–945. https://doi.org/10.1007/s10722-020-00893-9
Gengatharan A, Dykes GA, Choo WS (2015) Betalains: natural plant
pigments with potential application in functional foods. LWT-
Food Sci Technol 64:645–649. https://doi.org/10.1016/j.lwt.
2015.06.052
Havananda T, Luengwilai K (2019) Variation in floral antioxidant
activities and phytochemical properties among butterfly pea
(Clitoria ternatea L.) germplasm. Genet Resour Crop Eviron
66:645–658. https://doi.org/10.1007/s10722-018-00738-6
Herrera MC, De Castro ML (2005) Ultrasound-assisted extraction of
phenolic compounds from strawberries prior to liquid chromato-
graphic separation and photodiode array ultraviolet detection.
J Chromatogr A 1100:1–7. https://doi.org/10.1016/j.chroma.
2005.09.021
Iamsaard S, Burawat J, Kanla P et al (2014) Antioxidant activity and
protective effect of Clitoria ternatea flower extract on testicular
damage induced by ketoconazole in rats. J Zhejiang Univ Sci B
15:548–555. https://doi.org/10.1631/jzus.b1300299
Jain T (2009) Microwave assisted extraction for phytoconstituents: an
overview. Asian J Chem 2:19–25
Jamil N, Zairi MNM, Nasim NAIM et al (2018) Influences of
environmental conditions to phytoconstituents in Clitoria ter-natea (butterfly pea flower): a review. J Sci Technol 10:208–228
Kamilla L, Mnsor SM, Ramanathan S et al (2009) Antimicrobial
activity of Clitoria ternatea (L.) extracts. Pharmacologyonline
1:731–738
Kamkaen N, Wilkinson JM (2009) The antioxidant activity of Clitoriaternatea flower petal extracts and eye gel. Phytother Res
23:1624–1625. https://doi.org/10.1002/ptr.2832
Kazuma K, Noda N, Suzuki M (2003) Flavonoid composition related
to petal color in different lines of Clitoria ternatea. Phytochem-
istry 64:1133–1139. https://doi.org/10.1016/s0031-9422(03)
00504-1
Kosai P, Sirisidthi K, Jiraungkoorskul K et al (2015) Review on
ethnomedicinal uses of memory boosting herb, butterfly pea,
Clitoria ternatea. J Nat Remedies 15:71–76. https://doi.org/10.
18311/jnr/2015/480
J Food Sci Technol (June 2021) 58(6):2054–2067 2065
123
Page 13
Kumar BS, Bhat KI (2011) In-vitro cytotoxic activity studies of
Clitoria ternatea linn flower extracts. Int J Pharma Sci Rev Res
6:120–121
Lakshan SAT, Jayanath NY, Abeysekera WPKM et al (2019) A
commercial potential blue pea (Clitoria ternatea L.) flower
extract incorporated beverage having functional properties. Evid
Based Complement Altern Med 2019:1–13. https://doi.org/10.
1155/2019/2916914
Leong CR, Azizi K, Afif M et al (2017) Anthocyanins from Clitoriaternatea attenuate food-borne Penicillium expansum and its
potential application as food biopreservative. Nat Prod Sci
23:125–131. https://doi.org/10.20307/nps.2017.23.2.125
Lopez Prado AS, Shen Y, Ardoin R et al (2019) Effects of different
solvents on total phenolic and total anthocyanin contents of
Clitoria ternatea L. petal and their anti-cholesterol oxidation
capabilities. Int J Food Sci Technol 54:424–431. https://doi.org/
10.1111/ijfs.13953
Ludin AA, Al-Alwani MA, Mohamad AB et al (2018) Utilization of
natural dyes from Zingiber officinale leaves and Clitoria ternateaflowers to prepare new photosensitisers for dye-sensitised solar
cells. Int J Electrochem Sci 13(8):7451–7465. https://doi.org/10.
20964/2018.08.04
Mahmad N, Taha RM, Othman R et al (2018) Anthocyanin as
potential source for antimicrobial activity in Clitoria ternatea L.
and Dioscorea alata L. Pigm Resin Technol 47:490–495. https://
doi.org/10.1108/prt-11-2016-0109
Mathew N, Anitha MG, Bala TSL et al (2009) Larvicidal activity of
Saraca indica, Nyctanthes arbor-tristis, and Clitoria ternateaextracts against three mosquito vector species. Parasitol Res
104:1017–1025. https://doi.org/10.1007/s00436-008-1284-x
Mauludifia F, Astrinia SD, Meiranti KA et al (2019) Production of
natural colorant powder from Clitoria ternatea L. using tray
dryer which is dehumidified by zeolite. J Phys Conf Ser
1295:012018
Mehmood A, Ishaq M, Zhao L et al (2019) Impact of ultrasound and
conventional extraction techniques on bioactive compounds and
biological activities of blue butterfly pea flower (Clitoriaternatea L.). Ultrason Sonochem 51:12–19. https://doi.org/10.
1016/j.ultsonch.2018.10.013
Mukherjee PK, Kumar V, Kumar NS et al (2008) The Ayurvedic
medicine Clitoria ternatea-From traditional use to scientific
assessment. J Ethnopharmacol 120:291–301. https://doi.org/10.
1016/j.jep.2008.09.009
Nair V, Bang WY, Schreckinger E et al (2015) Protective role of
ternatin anthocyanins and quercetin glycosides from butterfly
pea (Clitoria ternatea Leguminosae) blue flower petals against
lipopolysaccharide (LPS)-induced inflammation in macrophage
cells. J Agric Food Chem 63:6355–6365. https://doi.org/10.
1021/acs.jafc.5b00928
Neda GD, Rabeta MS, Ong MT (2013) Chemical composition and
anti-proliferative properties of flowers of Clitoria ternatea. IntFood Res J 20:1229–1234
Nguyen GKT, Zhang S, Nguyen NTK et al (2011) Discovery and
characterization of novel cyclotides originated from chimeric
precursors consisting of albumin-1 chain a and cyclotide
domains in the Fabaceae family. J Biol Chem 286:24275–
24287. https://doi.org/10.1074/jbc.m111.229922
Nirmal SA, Bhalke RD, Jadhav RS et al (2008) Anthelmintic activity
of Clitoria ternatea. Pharmacologyonline 1:114–119
Nithianantham K, Ping KY, Latha LY et al (2013) Evaluation of
hepatoprotective effect of methanolic extract of Clitoria ternatea(Linn.) flower against acetaminophen-induced liver damage.
Asian Pac J Trop Dis 3:314–319. https://doi.org/10.1016/s2222-
1808(13)60075-4
Oguis GK, Gilding EK, Jackson MA et al (2019) Butterfly pea
(Clitoria ternatea), a cyclotide-bearing plant with applications in
agriculture and medicine. Front Plant Sci 10:645. https://doi.org/
10.3389/fpls.2019.00645
Perfect JR (2016) Is there an emerging need for new antifungals?
Expert Opin Emerg Drugs 21:129–131. https://doi.org/10.1517/
14728214.2016.1155554
Pham TN, Nguyen DC, Lam TD et al (2019) Extraction of
anthocyanins from Butterfly pea (Clitoria ternatea L. flowers)
in Southern Vietnam: response surface modeling for optimiza-
tion of the operation conditions. IOP Conf Ser Mater Sci Eng
542:012032. https://doi.org/10.1088/1757-899x/542/1/012032
Pham-Huy LA, He H, Pham-Huy C (2008) Free radicals, antioxidants
in disease and health. Int J Biomed Sci 4:89–96
Phrueksanan W, Yibchok-anun S, Adisakwattana S (2014) Protection
of Clitoria ternatea flower petal extract against free radical-
induced hemolysis and oxidative damage in canine erythrocytes.
Res Vet Sci 97:357–363. https://doi.org/10.1016/j.rvsc.2014.08.
010
Rabeta MS, An Nabil Z (2013) Total phenolic compounds and
scavenging activity in Clitoria ternatea and Vitex negundo linn.
Int Food Res J 20:495–500
Rajamanickam M, Kalaivanan P, Sivagnanam I (2015) Evaluation of
anti-oxidant and anti-diabetic activity of flower extract of
Clitoria ternatea L. J Appl Pharm Sci 5:131–138. https://doi.
org/10.7324/japs.2015.50820
Ranaganayaki S, Singh AK (1979) Isolation and identification of
pigments of the flowers of Clitoria ternatea. J Indian Chem Soc
56:1037–1038
Reid R, Sinclair DF (1980) An evaluation of a collection of Clitoria
ternatea for forage and grain production. CSIRO, Division of
Tropical Crops & Pastures; 1980. ISSN: 01596071
Ruenroengklin N, Zhong J, Duan X et al (2008) Effects of various
temperatures and pH values on the extraction yield of phenolics
from litchi fruit pericarp tissue and the antioxidant activity of the
extracted anthocyanins. Int J Mol 9:1333–1341. https://doi.org/
10.3390/ijms9071333
Saito N, Abe K, Honda T et al (1985) Acylated delphinidin glucosides
and flavonols from Clitoria ternatea. Phytochemistry 24:1583–
1586
Scheffler RJ, Colmer S, Tynan H et al (2013) Antimicrobials, drug
discovery, and genome mining. Appl Microbiol Biotechnol
97:969–978. https://doi.org/10.1007/s00253-012-4609-8
Senica M, Stampar F, Petkovsek MM (2019) Different extraction
processes affect the metabolites in blue honeysuckle (Loniceracaerulea L. subsp. edulis) food products. Turk J Agric For
43:576–585. https://doi.org/10.3906/tar-1907-48
Senkal BC, Uskutoglu T, Cesur C et al (2019) Determination of
essential oil components, mineral matter, and heavy metal
content of Salvia virgata Jacq. grown in culture conditions. Turk
J Agric For 43:395–404. https://doi.org/10.3906/tar-1812-84
Shen Y, Du L, Zeng H et al (2016) Butterfly pea (Clitoria ternatea)seed and petal extracts decreased Hep-2 carcinoma cell viability.
Int J Food Sci Technol 51:1860–1868. https://doi.org/10.1111/
ijfs.13158
Shyamkumar IB, Ishwar B (2012) Anti-inflammatory, analgesic and
phytochemical studies of Clitoria ternatea Linn flower extract.
Int Res J Pharm 3:208–210
Singh NK, Garabadu D, Sharma P et al (2018) Anti-allergy and anti-
tussive activity of Clitoria ternatea L. in experimental animals.
J Ethnopharmacol 224:15–26. https://doi.org/10.1016/j.jep.2018.
05.026
Keka S, Saha PD, Ramya V et al (2012) Improved extraction of
natural blue dye from butterfly pea using microwave assisted
methodology to reduce the effect of synthetic blue dye. Int J
Chem Technol 4:57–65. https://doi.org/10.3923/ijct.2012.57.65
Siti Azima AM, Noriham A, Manshoor N (2014) Anthocyanin
content in relation to the antioxidant activity and colour
2066 J Food Sci Technol (June 2021) 58(6):2054–2067
123
Page 14
properties of Garcinia mangostana peel, Syzigium cumini andClitoria ternatea extracts. Int Food Res J 21:2369–2375
Sivaranjan VV, Balachandran I (1994) Ayurvedic drugs and their
plant sources. Oxford & IBH Publishers Pvt., Ltd., New Delhi
Sreejith S, Samant MP, Jakhar JK et al (2014) Modeling the impact of
extraction conditions on functional properties of gelatin from
scales of blackspotted croaker (Protonibea diacanthus). Proc
Natl A Sci India B 84:1021–1029. https://doi.org/10.1007/
s40011-013-0259-6
Srichaikul B (2018) Ultrasonication extraction, bioactivity, antioxi-
dant activity, total flavonoid, total phenolic and antioxidant of
Clitoria ternatea linn flower extract for anti-aging drinks.
Pharmacogn Mag 14:322. https://doi.org/10.4103/pm.pm_206_
17
Subramanian MS, Prathyusha P (2011) Pharmaco-phytochemical
characterization of Clitoria ternatea Linn. Int J Pharmtech Res
3:606–612
Terahara N, Saito N, Honda T et al (1989) Structure of ternatin D1, an
acylated anthocyanin from Clitoria ternatea flowers. Tetrahe-
dron Lett 30:5305–5308. https://doi.org/10.1016/s0040-4039(01)
93771-2
Terahara N, Oda M, Matsui T et al (1996) Five new anthocyanins,
ternatins A3, B4, B3, B2, and D2, from Clitoria ternatea flowers.J Nat 59:139–144. https://doi.org/10.1021/np960050a
Terahara N, Toki K, Saito N et al (1998) Eight new anthocyanins,
ternatins C1–C5 and D3 and preternatins A3 and C4 from young
Clitoria ternatea flowers. J Nat 61:1361–1367. https://doi.org/10.1021/np980160c
Uma B, Prabhakar K, Rajendran S (2009) Phytochemical analysis and
antimicrobial activity of Clitorea ternatea Linn against extended
spectrum beta lactamase producing enteric and urinary patho-
gens. Asian J Pharm Clin Res 2:94–96
Wen C, Zhang J, Zhang H et al (2018) Advances in ultrasound
assisted extraction of bioactive compounds from cash crops: a
review. Ultrason Sonochem 48:538–549. https://doi.org/10.1016/
j.ultsonch.2018.07.018
Zakaria NNA, Okello EJ, Howes MJ et al (2018) In vitro protective
effects of an aqueous extract of Clitoria ternatea L. flower
against hydrogen peroxide-induced cytotoxicity and UV-induced
mtDNA damage in human keratinocytes. Phytother Res
32:1064–1072. https://doi.org/10.1002/ptr.6045
Zhang Z, Pang X, Ji Z et al (2001) Role of anthocyanin degradation in
litchi pericarp browning. Food Chem 75:217–221. https://doi.
org/10.1016/s0308-8146(01)00202-3
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
J Food Sci Technol (June 2021) 58(6):2054–2067 2067
123